scholarly journals Changes in levels of actin and tubulin mRNAs upon the lectin activation of lymphocytes.

1984 ◽  
Vol 4 (9) ◽  
pp. 1754-1760 ◽  
Author(s):  
E McCairns ◽  
D Fahey ◽  
G E Muscat ◽  
M Murray ◽  
P B Rowe
Keyword(s):  
Human Peripheral Blood ◽  
Mrna Level ◽  
Cdna Clones ◽  
Resting Cells ◽  
Beta Tubulin ◽  
Beta Actin ◽  
Rna Content ◽  
Mrna Ratio ◽  
Tubulin Mrna ◽  
Actin Mrna

The expression of beta-actin, gamma-actin, alpha-tubulin, and beta-tubulin mRNA during the lectin activation of human peripheral blood lymphocytes was examined with specific cDNA clones. The resting lymphocyte has a low level of both alpha- and beta-tubulin mRNAs, and these increase 10-fold after 72 h of lectin stimulation in which maximum cell transformation is achieved. Although there is a slight increase in tubulin mRNA during the first 6 h, most of the increase occurs between 6 and 24 h as the cells start to increase their RNA content and progress from G0 into G1. Both beta- and gamma-actin mRNAs are more abundant than the tubulin mRNAs in resting cells, with beta-actin mRNA being the major species. Upon activation, beta-actin mRNA increases threefold, whereas gamma-actin mRNA increases almost sixfold. Both beta- and gamma-actin mRNA are elevated 2.5-fold as early as 6 h, the gamma-actin mRNA level then increasing more than beta-actin between 6 and 24 h, resulting in the reduced beta-actin/gamma-actin mRNA ratio. The lectin-stimulated lymphocyte has a similar beta-actin/gamma-actin mRNA ratio as that of the human leukemic T-lymphoblast cell line CCRF-CEM. These increases are over and above the general increase in polyadenylated RNA content upon lectin activation. On returning to a noncycling state, the levels of these cytoskeletal mRNAs decrease. There were two beta-tubulin mRNAs present in lymphocyte cytoplasm, one of 1.8 kilobases and one of 2.8 kilobases in length. The nongrowing lymphocytes had relatively lower levels of the larger sized mRNA. Upon stimulation, the relative level of the larger mRNA was increased, and at 72 h the cells had approximately equal levels of both mRNAs as did the leukemic lymphoblasts.

Changes in levels of actin and tubulin mRNAs upon the lectin activation of lymphocytes

1984 ◽  
Vol 4 (9) ◽  
pp. 1754-1760
Author(s):  
E McCairns ◽  
D Fahey ◽  
G E Muscat ◽  
M Murray ◽  
P B Rowe
Keyword(s):  
Human Peripheral Blood ◽  
Mrna Level ◽  
Cdna Clones ◽  
Resting Cells ◽  
Beta Tubulin ◽  
Beta Actin ◽  
Rna Content ◽  
Mrna Ratio ◽  
Tubulin Mrna ◽  
Actin Mrna

The expression of beta-actin, gamma-actin, alpha-tubulin, and beta-tubulin mRNA during the lectin activation of human peripheral blood lymphocytes was examined with specific cDNA clones. The resting lymphocyte has a low level of both alpha- and beta-tubulin mRNAs, and these increase 10-fold after 72 h of lectin stimulation in which maximum cell transformation is achieved. Although there is a slight increase in tubulin mRNA during the first 6 h, most of the increase occurs between 6 and 24 h as the cells start to increase their RNA content and progress from G0 into G1. Both beta- and gamma-actin mRNAs are more abundant than the tubulin mRNAs in resting cells, with beta-actin mRNA being the major species. Upon activation, beta-actin mRNA increases threefold, whereas gamma-actin mRNA increases almost sixfold. Both beta- and gamma-actin mRNA are elevated 2.5-fold as early as 6 h, the gamma-actin mRNA level then increasing more than beta-actin between 6 and 24 h, resulting in the reduced beta-actin/gamma-actin mRNA ratio. The lectin-stimulated lymphocyte has a similar beta-actin/gamma-actin mRNA ratio as that of the human leukemic T-lymphoblast cell line CCRF-CEM. These increases are over and above the general increase in polyadenylated RNA content upon lectin activation. On returning to a noncycling state, the levels of these cytoskeletal mRNAs decrease. There were two beta-tubulin mRNAs present in lymphocyte cytoplasm, one of 1.8 kilobases and one of 2.8 kilobases in length. The nongrowing lymphocytes had relatively lower levels of the larger sized mRNA. Upon stimulation, the relative level of the larger mRNA was increased, and at 72 h the cells had approximately equal levels of both mRNAs as did the leukemic lymphoblasts.

Regulation of tubulin and actin mRNA production in rat brain: expression of a new beta-tubulin mRNA with development

1983 ◽  
Vol 3 (8) ◽  
pp. 1333-1342
Author(s):  
J F Bond ◽  
S R Farmer
Keyword(s):  
Rat Brain ◽  
Morphological Changes ◽  
In Vitro Translation ◽  
Brain Maturation ◽  
Cdna Clones ◽  
Mrna Levels ◽  
Beta Tubulin ◽  
Mrna Species ◽  
Tubulin Mrna ◽  
Actin Mrna

The expression of alpha-tubulin, beta-tubulin, and actin mRNA during rat brain development has been examined by using specific cDNA clones and in vitro translation techniques. During brain maturation (0 to 80 days postnatal), these mRNA species undergo a significant decrease in abundance. The kinetics of this decrease varies between the cerebrum and the cerebellum. These mRNAs are most abundant in both tissues during week 1 postnatal, each representing 10 to 15% of total mRNA activity. Both alpha- and beta-tubulin mRNA content decreases by 90 to 95% in the cerebrum after day 11 postnatal, and 70 to 80% decreases in the cerebellum after day 16. Actin sequences also decrease but to a lesser extent in both tissues (i.e., 50%). These decreases coincide with the major developmental morphological changes (i.e., neurite extension) occurring during this postnatal period. These studies have also identified the appearance of a new 2.5-kilobase beta-tubulin mRNA species, which is more predominant in the cerebellar cytoplasm. The appearance of this form occurs at a time when the major 1.8-kilobase beta-tubulin mRNA levels are declining. The possibility that the tubulin multigene family is phenotypically expressed and then this expression responds to the morphological state of the nerve cells is discussed.

scholarly journals Regulation of tubulin and actin mRNA production in rat brain: expression of a new beta-tubulin mRNA with development.

1983 ◽  
Vol 3 (8) ◽  
pp. 1333-1342 ◽  
Author(s):  
J F Bond ◽  
S R Farmer
Keyword(s):  
Rat Brain ◽  
Morphological Changes ◽  
In Vitro Translation ◽  
Brain Maturation ◽  
Cdna Clones ◽  
Mrna Levels ◽  
Beta Tubulin ◽  
Mrna Species ◽  
Tubulin Mrna ◽  
Actin Mrna

The expression of alpha-tubulin, beta-tubulin, and actin mRNA during rat brain development has been examined by using specific cDNA clones and in vitro translation techniques. During brain maturation (0 to 80 days postnatal), these mRNA species undergo a significant decrease in abundance. The kinetics of this decrease varies between the cerebrum and the cerebellum. These mRNAs are most abundant in both tissues during week 1 postnatal, each representing 10 to 15% of total mRNA activity. Both alpha- and beta-tubulin mRNA content decreases by 90 to 95% in the cerebrum after day 11 postnatal, and 70 to 80% decreases in the cerebellum after day 16. Actin sequences also decrease but to a lesser extent in both tissues (i.e., 50%). These decreases coincide with the major developmental morphological changes (i.e., neurite extension) occurring during this postnatal period. These studies have also identified the appearance of a new 2.5-kilobase beta-tubulin mRNA species, which is more predominant in the cerebellar cytoplasm. The appearance of this form occurs at a time when the major 1.8-kilobase beta-tubulin mRNA levels are declining. The possibility that the tubulin multigene family is phenotypically expressed and then this expression responds to the morphological state of the nerve cells is discussed.

Gestational changes in Ca2+ transport across rat placenta and mRNA for calbindin9K and Ca(2+)-ATPase

1992 ◽  
Vol 263 (4) ◽  
pp. R930-R935 ◽  
Author(s):  
J. D. Glazier ◽  
D. E. Atkinson ◽  
K. L. Thornburg ◽  
P. T. Sharpe ◽  
D. Edwards ◽  
...  
Keyword(s):  
Diffusion Coefficient ◽  
Calcium Transport ◽  
Hybridization Analysis ◽  
Northern Hybridization ◽  
Gestational Period ◽  
Beta Actin ◽  
Mrna Ratio ◽  
Calcium Clearance ◽  
Actin Mrna ◽  
Rate Limiting

The unidirectional maternofetal clearance (Kmf) of 45Ca was measured across the rat placenta over the last one-third of gestation. Kmf for 45Ca normalized to its diffusion coefficient in water (Kmf/Dw) increased 72-fold between days 15 and 22 of gestation from 3.5 +/- 0.3 to 253.1 +/- 22.0 cm/g placenta, respectively. At 15 and 18 days of gestation, Kmf/Dw for 45Ca was similar to Kmf/Dw for the paracellular marker [14C]mannitol, but at 21 and 22 days of gestation, Kmf/Dw for 45Ca was significantly higher than Kmf/Dw for [14C]mannitol, indicating that an additional route of transfer, other than diffusion, becomes available to calcium during this period. Northern hybridization analysis demonstrated that rat placental calbindin9K-to-beta-actin mRNA ratio increased 135-fold between 15 and 22 days of gestation and was temporally associated with the gestational increase in Kmf/Dw for 45Ca. In contrast, rat placental Ca(2+)-ATPase-to-beta-actin mRNA ratio increased only two- to threefold over the same gestational period and did not mirror the gestational changes in calcium clearance. These trends suggest that the expression of placental calbindin9K, but not Ca(2+)-ATPase, may be rate limiting to placental calcium transport in the rat.

scholarly journals Growth hormone regulation of the expression of differentiation-dependent genes in preadipocyte Ob1771 cells

1986 ◽  
Vol 238 (1) ◽  
pp. 123-129 ◽  
Author(s):  
A Doglio ◽  
C Dani ◽  
P Grimaldi ◽  
G Ailhaud
Keyword(s):  
Growth Hormone ◽  
Mrna Level ◽  
Protein A ◽  
Transcriptional Level ◽  
Hormone Regulation ◽  
Beta Actin ◽  
Adipose Conversion ◽  
Specific Mrnas ◽  
Actin Mrna ◽  
Glycerol 3 Phosphate Dehydrogenase

The adipose conversion of Ob1771 preadipocytes, during exposure to medium containing bovine serum and supplemented with growth hormone, is accompanied by the acquisition of phenotypic markers and the increased accumulation of specific mRNAs. The expression of lipoprotein lipase, and that of unidentified pOb24 and pGH3 mRNAs, are early events which are independent of growth hormone supplementation. By contrast, the late expression of mRNAs encoding for glycerol-3-phosphate dehydrogenase and p422 protein (a myelin-P2 homologue) and that of glycerol-3-phosphate dehydrogenase activity require the presence of growth hormone. The abundance of beta-actin mRNA does not change during differentiation. Runoff transcription by nuclei isolated from untreated or growth hormone-treated cells reveal little or no change in the rates of transcription of pOb24, pGH3 and beta-actin mRNAs. By contrast, the transcription rate of the p422 gene increases markedly (greater than 6-fold) in nuclei of growth hormone-treated cells. However, the p422 mRNA is more abundant than would be predicted by its nuclear transcription alone, suggesting, in Ob1771 cells exposed to growth hormone, that there is a post-transcriptional level of control. These results indicate that the permissive role of growth hormone during adipose cell differentiation is related to terminal events only and that its effects can be seen both at the protein and mRNA level. These results strongly suggest that an increased rate of specific transcription is primarily responsible for the accumulation of mRNAs during exposure to growth hormone.

Abstract 3465: Fkbp12.6 Overexpression in Mouse Cardiac Myocytes Offers Minor Protection Against Pressure Overload-Induced Cardiac Remodeling and Failure

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Laurent Vinet ◽  
Mylène Pezet ◽  
Miresta Prévilon ◽  
Barnabas Gellen ◽  
Celine Dachez ◽  
...  
Keyword(s):  
Heart Failure ◽  
Cardiac Remodeling ◽  
Protein Level ◽  
Mrna Level ◽  
Pressure Overload ◽  
Calcium Handling ◽  
Heart Catheterization ◽  
Mrna Ratio ◽  
Gravimetric Data ◽  
Actin Mrna

Alterations in RyR2 function is a hallmark of heart failure (HF). Decreased FKBP12.6 binding to RyR2 has been put forward to explain the diastolic SR Ca 2+ leakage observed in this condition. Previous work in the mouse has shown that cardiac FKBP12.6 overexpression protects against the development of myocardial infarction-induced heart failure. We tested the hypothesis that cardiac FKBP12.6 overexpression protects against transverse aortic constriction (TAC)-induced cardiac remodeling and failure. We used a mouse model of conditional cardiac-specific FKBP12.6 overexpression. Ten weeks after TAC, male transgenic mice (TG) and their wild-type controls (WT) underwent heart catheterization. Hemodynamic and gravimetric data are shown in the table . Ventricular expression of the hypertrophic gene program and calcium handling proteins were assessed by real-time PCR and Western blot, respectively. Ten weeks after TAC, the mortality rate was 23% in WT and 13% in TG (14/60 vs 5/39, ns). The percentage of mice with HF, estimated on the presence of pulmonary oedema, was 42% in WT-TAC and 32% in TG-TAC (15/36 vs 7/22, ns). BNP mRNA level increased 2.8 fold in WT-TAC (p<0.01 vs WT-Shams) and 2.4 fold in TG-TAC (p<0.01 vs TG-Shams). α-skeletal actin mRNA level increased 4.3 fold in WT-TAC (p<0.001 vs WT-Shams) and 3.8 fold in TG-TAC (p<0.001 vs TG-Shams). β-MHC/α-MHC mRNA ratio increased 2.8 fold in WT-TAC (p<0.01 vs WT-Shams) and 4.3 fold in TG-TAC (p<0.05 vs TG-Shams). RyR2 protein level decreased by 58% in WT-TAC and 41% in TG-TAC (p<0.01 and p<0.05 vs sham-operated mice, respectively). SERCA2a protein level decreased by 29% in WT-TAC and 16% in TG-TAC (p<0.01 and p<0.05 vs sham-operated mice, respectively). No statistical difference was found between TG-TAC and WT-TAC for any of these parameters. Conclusion: Cardiac FKBP 12.6 overexpression offers weak protection if any against TAC-induced cardiac remodeling and failure in the mouse.

Quantitation of intratumoral thymidylate synthase expression predicts for disseminated colorectal cancer response and resistance to protracted-infusion fluorouracil and weekly leucovorin.

1997 ◽  
Vol 15 (10) ◽  
pp. 3223-3229 ◽  
Author(s):  
C G Leichman ◽  
H J Lenz ◽  
L Leichman ◽  
K Danenberg ◽  
J Baranda ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Thymidylate Synthase ◽  
Mrna Level ◽  
Genetic Determinants ◽  
Response Data ◽  
Beta Actin ◽  
Actin Mrna ◽  
Colorectal Cancer Patients ◽  
Tumor Biopsies

PURPOSE Response rates to fluorouracil (5-FU)-based therapy remain low. As new, active agents are being tested, information regarding specific intratumoral genetic determinants of chemotherapy sensitivity or resistance can be used to plan therapy rationally. Intratumoral thymidylate synthase (TS) quantitation may be among the most important determinants of sensitivity or resistance to 5-FU. MATERIALS AND METHODS Forty-six disseminated colorectal cancer patients had measurable tumor biopsies for polymerase chain reaction (PCR)-based determination of TS mRNA pretreatment. Protracted infusion of 5-FU 200 mg/m2/d for 21 days with weekly intravenous leucovorin 20 mg/m2 each cycle was given. After two cycles, responses were evaluated. Response data were correlated with independently determined intratumoral ratios of TS/beta-actin mRNA for each patient. RESULTS TS/beta-actin ratios were successfully obtained for 42 patients (91%). TS/beta-actin ratios ranged from 0.3 x 10(-3) to 18.2 x 10(-3) (median, 3.5 x 10[-3]). Twelve patients (26%) responded to treatment (median TS/beta-actin ratio, 1.7 x 10[+3]). Thirty-four patients did not respond (median TS/beta-actin ratio, 5.6 x 10[-3]). No patient with a TS mRNA level greater than 4.1 x 10(-3) responded. The median TS/beta-actin ratio (3.5 x 10[-3]) significantly segregated responders from nonresponders (P = .001). Median survival for patients with TS/beta-actin ratios < or = 3.5 x 10(-3) was 13.6 months; for patients with TS/beta-actin ratios greater than 3.5 x 10(-3), it was 8.2 months (P = .02). CONCLUSION For this cohort, the intratumoral TS/beta-actin ratio had a statistically significant association with response and survival. This relationship for other 5-FU schedules remains unknown. Confirmation of these data in a larger patient population could lead to determination of therapy for disseminated colorectal cancer based on a specific intratumoral molecular parameter.

Determination of skeletal muscle calpain and calpastatin activities during maturation

1991 ◽  
Vol 261 (6) ◽  
pp. E677-E683 ◽  
Author(s):  
B. R. Ou ◽  
N. E. Forsberg
Keyword(s):  
Skeletal Muscle ◽  
Muscle Protein ◽  
Mrna Level ◽  
Beta Tubulin ◽  
Mrna Concentration ◽  
Beta Actin ◽  
Age Related ◽  
Synthetic Capacity ◽  
Calpain Ii ◽  
Age Related Changes

Our objectives were to characterize events underlying changes in skeletal muscle calpain and calpastatin activities, using maturation as a model. Muscle samples were taken from rabbits of four ages (newborn and 1, 2, and 5 mo old). Concentrations of RNA and protein and activities of calpains I and II and calpastatin were determined. Steady-state concentrations of mRNAs encoding calpain I, calpain II, calpastatin, alpha- and beta-tubulin, and beta-actin were determined using Northern blot analysis. Calpain and calpastatin activities declined markedly between birth and 1 mo of age and remained unchanged thereafter. Several factors accounted for the neonatal losses of calpains and calpastatin. First, muscle protein concentration increased between birth and 1 mo of age and diluted calpain and calpastatin specific activities. Second, there was a marked reduction of muscle RNA concentration between birth and 1 mo of age, which indicates that protein synthetic capacity declined with age. Finally, calpastatin mRNA concentration declined between birth and 1 mo of age and further contributed to developmental losses of calpastatin activity. Calpain I mRNA concentration was unaffected by age, and although calpain II mRNA concentration declined with age, losses were not detected between birth and 1 mo; hence age-related changes in calpain I and II activities are not mediated at the mRNA level. The age-related reductions in calpain II and calpastatin mRNA concentrations resembled age-related changes in alpha- and beta-tubulin and beta-actin mRNA concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Cell-cycle-specific and serum-dependent expression of gamma-actin mRNA in Swiss mouse 3T3 cells.

1988 ◽  
Vol 8 (6) ◽  
pp. 2288-2294 ◽  
Author(s):  
A S Masibay ◽  
P K Qasba ◽  
D N Sengupta ◽  
G P Damewood ◽  
T Sreevalsan
Keyword(s):  
Growth Factor ◽  
Calcium Ionophore ◽  
Swiss Mouse ◽  
Cdna Clones ◽  
Ionophore A23187 ◽  
Resting Cells ◽  
Mrna Levels ◽  
3T3 Cells ◽  
Specific Transcript ◽  
Actin Mrna

We isolated cDNA clones that represent genes whose expression is enhanced when resting Swiss mouse 3T3 cells are stimulated to proliferate with serum. Two clones (designated pME1 and pMR6) were analyzed further. A partial sequence analysis of the pME1 insert DNA indicated that it contained a 104-base-pair stretch with extensive homology to the 3' untranslated region of gamma actin. Similar analysis of the insert DNA from the pMR6 clone indicated that it did not correspond to any previously reported gene sequence. We used the pME1 clone as a probe to determine the level of gamma actin-specific transcript in 3T3 cells under a variety of conditions. The level of gamma actin-specific mRNA began to increase in resting cells upon serum stimulation and reached a peak at 6 h. Thereafter its level declined, and by 24 h it was hardly detectable. In contrast, pMR6-specific transcript was detectable in resting cells but remained elevated even at 24 h poststimulation. The level of gamma-actin mRNA was elevated in resting cells by 12-O-tetradecanoylphorbol-13-acetate, calcium ionophore A23187, and bombesin and to a lesser extent by cholera toxin, fibroblast-derived growth factor, and dibutyryl cyclic AMP. However, insulin, vasopressin, or epidermal growth factor failed to enhance gamma-actin mRNA levels in resting cells. Inhibitors of transcription diminished the induction of gamma-actin mRNA. Gamma-actin gene was superinduced in serum-stimulated cells by cycloheximide, an inhibitor of translation. Analysis of proteins from serum-stimulated cells by two-dimensional gel electrophoresis indicated that enhanced transcription of gamma-actin mRNA resulted in a concomitant increase in the corresponding actin protein. The possible role of gamma actin, a component of the cytoskeleton, in the regulation of cell growth is discussed.

Cold-induced alterations in uncoupling protein and its mRNA are seasonally dependent in ground squirrels

1995 ◽  
Vol 269 (2) ◽  
pp. R357-R364
Author(s):  
S. E. Nizielski ◽  
C. J. Billington ◽  
A. S. Levine
Keyword(s):  
Cold Exposure ◽  
Uncoupling Protein ◽  
Late Summer ◽  
Ground Squirrels ◽  
Mitochondrial Uncoupling ◽  
Mitochondrial Uncoupling Protein ◽  
Beta Actin ◽  
Mrna Ratio ◽  
Brown Adipose ◽  
Actin Mrna

We were interested in determining whether season affects the ability of cold exposure to increase brown adipose tissue (BAT) thermogenic function in 13-lined ground squirrels after acute and chronic cold (4 degrees C) exposure. Tissues were collected from animals in April and September after cold exposure for 12, 24, or 48 h. Animals chronically exposed to the cold (10 days) were killed in early May and mid-August. We found that mitochondrial uncoupling protein (UCP) concentrations varied seasonally, with concentrations in control animals (at 23 degrees C) higher in late summer (mid-August and September) than in the spring (April and early May). Cold exposure in late summer did not induce further increases in UCP concentrations. In contrast, when animals were cold exposed in the spring, UCP concentrations and total UCP increased. Surprisingly, 10 days at 4 degrees C did not cause a greater increase in UCP concentrations than did 24 h at 4 degrees C. Chronic cold exposure increased the UCP mRNA-to-beta-actin mRNA ratio 48% in May, whereas a fivefold increase occurred in August. GDP binding was increased after 12 h at 4 degrees C in April; in contrast, animals attempted to hibernate when placed in the cold in September, and no increase in GDP binding was observed. Chronic cold exposure caused GDP binding to increase at both times. These results indicate that mitochondrial UCP concentrations are seasonally regulated in the 13-lined ground squirrel.

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